香青兰总黄酮对RAW264.7巨噬细胞泡沫化及炎症反应的调控作用  

Regulation of Dracocephalum Moldavica total flavonoids on foaming and inflammatory response of RAW264.7 macrophages

作　　者：杜春妍 赵云丽 阮徐莉 王新春[2] 曹文疆[2] DU Chun-yan;ZHAO Yun-li;RUAN Xu-li;WANG Xin-chun;CAO Wen-jiang(College of Pharmacy,Shihezi University,Shihezi Xinjiang 832008,China;Dept of Pharmacy,the First Affiliated Hospital of the Medical College,Shihezi University,Shihezi Xinjiang 832008,China)

机构地区：[1]石河子大学药学院,新疆石河子832008 [2]石河子大学医学院第一附属医院药学部,新疆石河子832008

出　　处：《中国药理学通报》2023年第10期1973-1979,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81960766);兵团财政科技计划项目(No 2022AB020)。

摘　　要：目的研究香青兰总黄酮(total flavonoids of Dracocephalum moldavica L.,TFDM)对氧化低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导的小鼠单核巨噬细胞白血病细胞(RAW264.7)泡沫化及炎症的影响,进一步阐明TFDM抗动脉粥样硬化(atherosclerosis,AS)的作用机制。方法体外培养RAW264.7巨噬细胞,采用ox-LDL刺激诱导使其成为泡沫细胞,TFDM(25、50、100 mg·L^(-1))及辛伐他汀(10μmol·L^(-1))进行干预,油红O染色法观察胞内脂滴的聚集情况,CCK-8法检测细胞活力,活性氧试剂盒测定ROS的生成,实时荧光定量PCR测定细胞中NF-κB、NLRP3、caspase-1、IL-18和IL-1βmRNA的表达,免疫蛋白印迹法检测巨噬细胞中IκBα、NF-κB p65、NLRP3、pro-caspase-1、caspase-1、IL-1β以及IL-18蛋白的表达,ELISA法检测TNF-α和IL-10的表达。结果TFDM可以减少泡沫巨噬细胞的形成,降低炎症因子IL-1β、IL-18和TNF-α的表达,增加抑炎因子IL-10的表达;并且下调NF-κB p65、NLRP3、pro-caspase-1、caspase-1蛋白的表达,上调IκBα的蛋白表达。结论TFDM能够减轻巨噬细胞的泡沫化,抑制炎症因子的表达,从而可能延缓动脉粥样硬化的发展进程。其作用机制可能是通过抑制NF-κB途径,减少ox-LDL诱导的RAW264.7细胞中炎症介质的产生。Aim To investigate the effects of Dracocephalum Moldavica total flavonoids(TFDM)on the foaming lipoprotein(ox-LDL),and to further elucidate the mechanism of anti-atherosclerosis(AS)of TFDM.Methods RAW264.7 macrophages were cultured in vitro and induced to become foam cells by ox-LDL stimulation,and inflammation of mouse monocyte macrophage leukemia cells(RAW264.7)induced by oxidized low density TFDM(25,50,100 mg·L^(-1))and simvastatin(10μmol·L^(-1))were used for intervention.The accumulation of intracellular lipid droplets was observed by oil red O staining.Cell viability was determined by CCK-8 method,ROS production was determined by reactive oxygen species kit,and mRNA expressions of NF-κB,NLRP3,caspase-1,IL-18 and IL-1βwere determined by real-time PCR assay.The expressions of IκBα,NF-κB p65,NLRP3,pro-caspase-1,caspase-1,IL-1βand IL-18 protein in macrophages were detected by immunoblotting,and the expressions of TNF-αand IL-10 were detected by ELISA.Results TFDM could reduce the formation of foam macrophages,reduce the expressions of inflammatory cytokines IL-1β,IL-18 and TNF-α,and increase the expression of anti-inflammatory factor IL-10.The expressions of NF-κB p65,NLRP3,pro-caspase-1 and caspase-1 were down-regulated,and the expressions of IκBαwere up-regulated.Conclusions TFDM can reduce the foaming of macrophages and inhibit the expression of inflammatory factors,which may delay the development of atherosclerosis.The possible mechanism of action involves the reduction of the production of inflammatory mediators in ox-LDL-induced RAW264.7 cells by inhibiting NF-κB pathway.

关 键 词：香青兰总黄酮 巨噬细胞 泡沫细胞 炎症反应 NF-ΚB 动脉粥样硬化 

分 类 号：R-332[医药卫生] R284.1R329.24R364.5R543.5